QIAquick Nucleotide Removal Kit

For up to 10 μg oligonucleotide (17–40mers) and DNA (40 bp to 10 kb) cleanup from enzymatic reactions

Up to 95% recovery of ready-to-use DNA

Fast and convenient procedure

Cleanup of DNA up to 10 kb in three easy steps

Gel loading dye for convenient sample analysis

The QIAquick Nucleotide Removal Kit provides spin columns, buffers, and collection tubes for silica-membrane-based purification of oligonucleotides and DNA. Unincorporated nucleotides, salts, and other contaminants are removed and oligonucleotides (>17 nt) and DNA fragments ranging from 40 bp to 10 kb are purified using a simple and fast bind-wash-elute procedure and an elution volume of 30–200 μl. The procedure is fully automated on theQIAcube.

The QIAquick system uses a simple bind–wash–elute procedure (see flowchart "QIAquick Nucleotide Removal procedure"). Binding buffer is added directly to the sample, and the mixture is applied to the QIAquick spin column. Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

Handling

QIAquick spin columns are designed to provide two convenient handling options. The spin columns fit into a conventional table-top microcentrifuge. The QIAquick Nucleotide Removal Kit, in addition to other QIAGEN spin-column-based kits, can be fully automated on the QIAcube, enabling increased productivity and standardization of results (see figures "Spin column handling options A, and B").

Applications

DNA fragments purified with the QIAquick system are ready for direct use in all applications, including sequencing, microarray analysis, ligation and transformation, restriction digestion, labeling, and microinjection.

QIAquick® Spin Columns can now be used on any vacuum manifold with luer connectors, for example, the QIAvac 6S or QIAvac 24 with QIAvac Luer Adapters. This protocol is designed for removal of primers <10 bases, enzymes, salts, and unincorporated nucleotides from biotin-, or DIG-labeled DNA fragments and oligonucleotides >17 nucleotides.